This SBIR Phase I project will determine the feasibility of using Self-Digitization (SD) microfluidic technology as a commercially competitive nucleic acid quantification (NAQ) platform. Quantification of nucleic acids (both DNA and RNA) is utilized in various fields including general research, biomedical research, and clinical diagnostics, with specific applications in areas such as measuring gene expression levels, disease diagnostics including pathogen/viral load monitoring, cancer diagnostics, and more. Self-Digitization is a simple, robust technology that efficiently uses a network of channels and wells to spontaneously partition samples into an array of predefined well volumes. This platform is ideally suited for digital PCR (dPCR) applications. Digital PCR works by partitioning samples into thousands or even millions of individual volumes where each volume may or may not contain target DNA. Only volumes with target DNA give a positive signal resulting in a digital yes/no signal, from which Poisson statistics can then give direct determination of sample concentrations. Digital PCR directly provides absolute quantification, is robust against variations in reaction efficiency, is incredibly sensitive and has nearly unlimited resolution capabilities, making it technically superior to the current NAQ gold standard of qPCR. However, no commercial dPCR system is currently able to be economically competitive with qPCR. The objective of this proposal is to show the technical feasibility of a new centrifugal SD filling method within an optical disc (OD) style platform. The simplicity and flexibility of the SD system, its implementation within a method that enables parallelization and high-throughput experimentation, and the adaptation of existing commercialized components to achieve a complete platform will result in a cost-competitive product that will surpass existing commercial dPCR platforms, while also directly competing with qPCR systems. The approach will first validate the technical aspects of the platform by developing a thorough understanding of the loading/digitization mechanism, optimizing design features to maximize performance within a closed system, and to develop complete designs to meet needs in specific application categories. This will initially be carried out in scaled down versions to maximize iteration efficiency and then expanded to full prototype scale. Validation of this approach would then lead to future production of a mass producible device product and complete instrumentation setup. The product would be immediately useful for general biomedical research applications, and the workflow and nature of the closed system would also make it amenable to future clinical applications in a variety of medical fields.